JP6203598B2 - Sotron Smell Suppressor - Google Patents

Description

  The present invention relates to an inhibitor of odor caused by sotron.

  There are many malodorous molecules with different polarities and molecular weights in our living environment. Various deodorizing methods have been developed so far to deodorize various malodorous molecules. In general, deodorization methods are roughly classified into biological methods, chemical methods, physical methods, and sensory methods. Among the malodorous molecules, highly polar short chain fatty acids and amines can be deodorized by a chemical method, that is, neutralization reaction. In addition, sulfur compounds such as thiols can be deodorized by a physical method, that is, adsorption treatment. However, many malodorous molecules that cannot be handled by the conventional deodorization method remain. In addition, the deodorization method by adsorption treatment has a problem that bad odor is likely to be re-released. Furthermore, in these conventional methods, odors other than the target malodor may be erased. There is a need for a deodorizing method that can overcome these problems.

  There is also known a method of deodorizing bad odors by using a fragrance to recognize another odor more strongly. However, this method may cause discomfort with the smell of the fragrance. Furthermore, in order to mask malodor with other odors such as perfumes and fragrances, it is necessary to search for odorous substances that have an effective deodorizing action against the target malodorous substances. Traditionally, odor evaluation has been performed by sensory tests by experts. However, sensory tests have problems such as the need to train experts who can evaluate odors and low throughput. Therefore, conventionally, it has not been easy to search for an odorous substance exhibiting a deodorizing action.

  In mammals such as humans, the olfactory sensation is that an odorant molecule binds to an olfactory receptor on the olfactory nerve cell located in the olfactory epithelium in the upper nasal cavity, and the response of the receptor is transmitted to the central nervous system. It is recognized by. In humans, it is reported that there are about 400 olfactory receptors, and the genes encoding them are about 3% of all human genes. In general, olfactory receptors and odor molecules are associated with each other in a plurality of combinations. That is, individual olfactory receptors can accept multiple odor molecules with similar structures with different affinities, while individual odor molecules can be received by multiple olfactory receptors. Furthermore, it has been reported that an odor molecule that activates one olfactory receptor acts as an antagonist that inhibits activation of another olfactory receptor. The combination of these multiple olfactory receptor responses results in the recognition of individual odors.

  Therefore, even when the same odor molecule is present, if another odor molecule is present at the same time, the receptor response is inhibited by the other odor molecule, and the finally recognized odor may be completely different. This mechanism is called olfactory receptor antagonism. This odor modulation by receptor antagonism, unlike the deodorization method by adding another odor such as perfume or fragrance, can specifically lose the perception of malodor and Since there is no discomfort due to the smell, it is a preferred deodorizing means.

  Skin tanning agents (also called self-tanning agents or sunless tanning agents) are skin cosmetics that color the skin. As a component that changes the skin to brown, dihydroxyacetone (DHA) is mainly used alone or in combination with erythrulose. These ingredients react with the upper layers of the skin and color the skin brown. This coloring is thought to proceed by a browning reaction, but the details of the reaction mechanism have not been elucidated. The browning reaction is also called Maillard reaction in the field of food chemistry, and is a term indicating a reaction in which a nitrogen-containing compound such as amino acid or protein and a reducing sugar are polymerized to produce a brown polymer called melanoidin. The Maillard reaction is involved in the coloration of food and the generation of aroma components that occur by heating the food.

  3-Hydroxy-4,5-dimethyl-2 (5H) -furanone (Sotron) is expressed as “curry-like flavor”, “caramel incense”, “burnt sugar”, “burnt”, “spicy”, etc. It is a substance known to have an odor (Non-Patent Documents 1 to 4). Sotron is generally an off-flavor with a herbal unpleasant odor at high concentrations, but becomes molasses-like aroma at low concentrations. Sotron has been reported to induce the response of the olfactory receptor OR8D1 (Non-patent Document 5).

Synthetic perfume Chemistry and product knowledge, 2005, Chemical Industry Daily ACS Symp Ser., 2002, 836: 108-123 Journal of Biotechnology, 2006, 84 (10): 417 J. Agric. Food Chem., 1999, 47 (8): 3288-3291 PLoS Genet, 2012, 8 (7): e1002821

  When using commercially available skin tanning agents (self-tanning agents or sunless tanning agents), problems have been reported that are accompanied by a unique unpleasant odor expressed as earthy odor, burnt sugar, etc. (D. M. Hindenlang and M. E. McDonnell, Cosmetics & Toiletries magazine, 2008, Vol. 123, No. 7, p67-74), improvements were sought. When the cause of the unpleasant odor was investigated, 3-hydroxy-4,5-dimethyl-2 (5H) -furanone (Sotron) produced from dihydroxyacetone (DHA) contained in the skin tanning agent via methylglyoxal. Was found to be one of the causative substances. In order to suppress such an unpleasant odor caused by Sotron, it is required to control the odor of Sotron.

  On the other hand, Sotron has a molasses-like fragrance and can be used as a fragrance for foods and drinks. However, if the smell is too strong, it acts as an off-flavor. In order to adjust the odor of Sotron to an appropriate intensity and to make the aroma of food and drink more preferable, it is required to control the odor of Sotron.

  The present inventor has found that a substance that suppresses the response of the olfactory receptor OR8D1, which is known to respond to sotron, can suppress odor recognition by sotron by masking with olfactory receptor antagonism. .

Therefore, the present invention provides an inhibitor of odor caused by sotron, comprising at least one selected from the group consisting of the compounds shown in Table 1 below as an active ingredient.

  According to the odor inhibitor by sotron provided by the present invention, the odor by sotron can be selectively deodorized by masking by olfactory receptor antagonism. Therefore, the odor suppressor by Sotron provided by the present invention does not cause problems such as discomfort based on the odor of the fragrance that has occurred in the conventional deodorizing method using the odor eliminating agent or fragrance, For example, an unpleasant odor generated when using a conventional self-tanning agent (also referred to as a sunless tanning agent) can be eliminated.

Response of olfactory receptors to sotron. The horizontal axis represents individual olfactory receptors, and the vertical axis represents response intensity. Response of the olfactory receptor OR8D1 to various concentrations of sotron. n = 3, error bar = ± SE. Sensory evaluation of the sotron odor control effect of various compounds. n = 3, error bar = ± SE.

  In this specification, the term “masking” regarding odor refers to all means for making the target odor unrecognized or weakening recognition. “Masking” may include chemical means, physical means, biological means, and sensory means. For example, masking includes any means for removing odor molecules that cause the desired odor from the environment (eg, adsorption and chemical degradation of the odor molecules), to prevent the desired odor from being released into the environment. (For example, containment), a method of adding another odor such as a fragrance or a fragrance to make it difficult to recognize the target odor, and the like.

  In the present specification, “masking by olfactory receptor antagonism” is a form of the above-mentioned “masking” in a broad sense, and by applying the odor molecule of interest and other odor molecules together, This is a means for inhibiting the receptor response to the target odor molecule by the odor molecule and consequently changing the odor recognized by the individual. Masking by olfactory receptor antagonism is also distinguished from means for canceling the target odor by another strong odor, such as a fragrance, even if the means uses other odor molecules. An example of masking by olfactory receptor antagonism is the case of using a substance that inhibits the response of the olfactory receptor, such as an antagonist (antagonist). Applying a substance that inhibits the response to the receptor of an odor molecule that causes a specific odor suppresses the response of the receptor to the odor molecule, and consequently changes the odor perceived by the individual. Can do.

  As used herein, “scent by sotron” may be an odor caused by sotron (3-hydroxy-4,5-dimethyl-5 (2H) -furanone). As used herein, “scent by sotron” can typically be expressed as caramel flavor, burnt sugar scent, curry-like flavor, or the like. Further, in the present specification, “odor by sotron” means an unpleasant odor generated when a conventional self-tanning agent is applied to the skin, more specifically, a skin tanning agent containing dihydroxyacetone (DHA) is applied to the skin. It can be an unpleasant odor expressed as “Burnt sugar” that occurs when applied.

  As shown in FIG. 1, the present inventor has identified the olfactory receptor OR8D1 among the many olfactory receptors as the only receptor responsive to sotron. The olfactory receptor OR8D1 is an olfactory receptor expressed in human olfactory cells, and is registered with GenBank as GI: 50892781. OR8D1 is a protein consisting of an amino acid sequence represented by SEQ ID NO: 2 encoded by a gene having a base sequence represented by SEQ ID NO: 1.

  As shown in FIG. 2, OR8D1 responds to sotron in a concentration-dependent manner. Therefore, a substance that suppresses the response of OR8D1 causes a change in recognition of odor by sotron in the center of an individual by masking based on olfactory receptor antagonism, and as a result, selectively suppresses odor by sotron. it can.

  Therefore, in one aspect, the present invention provides a sonotron odor suppressor comprising a substance that suppresses the response of the olfactory receptor OR8D1 as an active ingredient. The sonotron odor suppressor of the present invention acts as an antagonist (antagonist) of OR8D1. In other words, the sonotron odor suppressor of the present invention comprises an OR8D1 antagonist as an active ingredient. Therefore, the odor suppressor by sotron of the present invention can change the recognition of odor by sotron in the center of an individual by masking based on olfactory receptor antagonism, and as a result, can suppress the odor by sotron. .

The active ingredient of the sotron odor suppressor according to the present invention includes the following compounds:
1- (2,3,4,7,8,8a-hexahydro-3,6,8,8-tetramethyl-1H-3a, 7-methanoazulen-5-yl) -ethanone (acetyl cedrene);
Oxacyclohexadecen-2-one (Habanolide®);
Citral;
ω-6-hexadecenelactone (ambretlide);
2-benzylideneheptanal (amylcinnamic aldehyde);
3- (4-tert-butylphenyl) propanal (Burgeonal);
(5E) -3-methylcyclopentadec-5-en-1-one (mussenon® delta);
1- (5,6,7,8-tetrahydro-3,5,5,6,8,8-hexamethyl-2-naphthalenyl) -ethanone (Tonalide®);
7-acetyl-1,2,3,4,5,6,7,8-octahydro-1,1,6,7-tetramethyl-naphthalene (iso-e super);
Muscon.

  The compound is an OR8D1 antagonist. The OR8D1 antagonist can inhibit the response of the olfactory receptor OR8D1 to sotron by 40% or more, preferably 50% or more, more preferably 75% or more, compared to the case where the antagonist is not present. In other words, the response of the olfactory receptor OR8D1 to sotron in the presence of the antagonist is reduced to 60% or less, preferably 50% or less, more preferably 25% or less of the response of the receptor in the absence of the antagonist. .

  Among the above OR8D1 antagonists, 1- (2,3,4,7,8,8a-hexahydro-3,6,8,8-tetramethyl-1H is preferable as an active ingredient of an inhibitor of odor caused by sotron. -3a, 7-methanoazulen-5-yl) -ethanone (acetyl cedrene).

  The sonotron-suppressing agent of the present invention contains at least one compound selected from the group consisting of the above-mentioned OR8D1 antagonists as an active ingredient. For example, the active ingredient of the sonotron odor suppressor of the present invention may be at least one selected from the group consisting of the above-mentioned OR8D1 antagonists, for example, either alone or in combination of any two or more. Preferably, the sotron odor suppressor of the present invention consists essentially of at least one selected from the group consisting of the above OR8D1 antagonists.

  As shown in Table 3, the OR8D1 antagonist suppresses the response of the olfactory receptor OR8D1 to sotron. The compound has been known as a fragrance material until now, but it has not been known that it has a function of antagonizing the response of an olfactory receptor to sotron and selectively suppressing odor caused by sotron.

  All of the above-mentioned OR8D1 antagonists can be purchased commercially (refer to "Synthetic Fragrance Chemistry and Product Knowledge" augmented and revised edition, Motoichi Into, Kagaku Kogyo Nippo, published in March 2005) . For example, it can be obtained from Firmenich S.A., Givaudan S.A., International Flavors & Fragrances Inc., Takasago International Corporation, Kao Corporation.

  In another aspect of the present invention, the OR8D1 antagonists listed above have a sotron scent, such as caramel flavor, burnt sugar scent, or curry-like flavor, or when applying a self-tanning agent to the skin or dihydroxyacetone. It can be used as an active ingredient for suppressing an unpleasant odor (for example, a burnt smell of sugar) generated when a product containing (DHA) is used.

  In one embodiment, the above-mentioned OR8D1 antagonist can be contained in a compound or composition for suppressing odor caused by sotron as an active ingredient for suppressing odor caused by sotron. Alternatively, it can be used for the production of an odor suppressor by Sotron, or for the production of a compound or composition for suppressing odor by Sotron.

  In one embodiment, the above-listed OR8D1 antagonists inhibit odor suppression by sotron in any compound or composition where odor suppression by sotron is desired, or in any environment where odor suppression by sotron is desired. Can be used as an active ingredient. Alternatively, the OR8D1 antagonist can be used as an active ingredient for the suppression of odor by sotron, for the production of a compound or composition in which the suppression of odor by sotron is desired. Examples of compounds or compositions where suppression of odor by sotron is desired include skin tanning agents (also called self-tanning agents or sunless tanning agents), for example skin tanning containing dihydroxyacetone (DHA) as a color former And other skin tanning agents that take advantage of the brown color reaction and other DHA containing products. In addition, Sotron is an aroma component contained in food and drink, but when the amount increases, it may work as an off-flavor. Therefore, another example of a compound or composition for which suppression of odor by sotron is desired includes foods and drinks for which it is desired to reduce off-flavor due to a high concentration of sotron and compositions containing them.

  In addition to the OR8D1 antagonists listed above, the above-described odor suppressor for odor caused by sotron and the compound or composition for suppressing odor caused by sotron appropriately contain other components having deodorizing effect depending on the purpose. You may do it. As the other components having the deodorizing effect, any known deodorant having a chemical or physical deodorizing effect can be used. For example, a deodorizing active ingredient extracted from a plant (for example, green tea) Extracts), organic acids, various amino acids and their salts, glyoxal, oxidizing agents, flavonoids, catechins, polyphenols, porous materials such as activated carbon and zeolite, inclusion agents such as cyclodextrins, photocatalysts, various masking Agents, etc. Further, the inhibitor and the compound or composition for suppressing odor caused by Sotron may contain any component that is generally added to deodorants and deodorants.

  As a further aspect, the present invention provides a method for suppressing odor caused by sotron, which comprises the step of allowing the odor caused by sotron to coexist with the above-mentioned antagonists of OR8D1.

  In one embodiment, the method requires suppression of odor recognition by sotrons by masking with olfactory receptor antagonism, preferably in the presence of odor by sotrons. The individual is applied with at least one selected from the group consisting of the above-mentioned OR8D1 antagonists so that the antagonists coexist with the sotron odor. As a result, OR8D1 and the antagonist bind to each other, and the response of OR8D1 to sotron is suppressed. Therefore, masking by olfactory receptor antagonism occurs, and the odor caused by sotron is suppressed. Preferred OR8D1 antagonists in the method are the same as those of the above-described odor suppressor by sotron.

  In the method of the present invention, the individual is not particularly limited as long as it is a mammal, but preferably a human. More specifically, in the method of the present invention, as an example of an individual who needs to suppress the recognition of odor by sotron, a skin tanning agent (or a self-tanning agent or a sunless tanning agent), more specifically, for example, Sotron from humans using skin tanning agents containing dihydroxyacetone (DHA) or other skin tanning agents utilizing brown reaction, humans using other DHA-containing products, and food and beverages Examples include humans who desire to reduce the above.

  In one embodiment of the method of the invention, the OR8D1 antagonist is formulated in a compound or composition containing sotron. As a result, the response of the olfactory receptor to sotron contained in the compound or composition is reduced, and the recognition of odor by sotron in the individual is suppressed, and as a result, the odor by sotron is suppressed. In yet another embodiment, the OR8D1 antagonist is formulated in a compound or composition that may generate sotron. This reduces the response of the olfactory receptor to sotron generated from the compound or composition, suppresses odor recognition by sotron in the individual, and consequently suppresses odor by sotron. Examples of compounds or compositions that may produce sotrons include skin tanning agents, such as skin tanning agents containing dihydroxyacetone (DHA) as a color former, other skin tanning agents that utilize a brown reaction, and Other products containing DHA are listed.

  As exemplary embodiments of the present invention, the following compositions, production methods, uses or methods are further disclosed herein. However, the present invention is not limited to these embodiments.

<1> An inhibitor of odor caused by sotron, comprising at least one selected from the group consisting of the compounds described in Table 1 as an active ingredient.

<2> Use of at least one selected from the group consisting of the compounds described in Table 1 for the suppression of odor by sotron.

<3> Use of at least one selected from the group consisting of the compounds described in Table 1 for the production of an odor suppressor by Sotron.

<4> At least one compound selected from the group consisting of the compounds described in Table 1 for use in suppressing odor caused by sotron.

<5> A method for suppressing odor caused by sotron, which comprises coexisting odor caused by sotron and at least one selected from the group consisting of the compounds described in Table 1 above.

<6> In the above items <1> to <5>, preferably, at least one selected from the group consisting of the compounds described in Table 1 is 1- (2,3,4,7,8,8a-hexahydro -3,6,8,8-tetramethyl-1H-3a, 7-methanoazulen-5-yl) -ethanone.

  EXAMPLES Hereinafter, an Example is shown and this invention is demonstrated more concretely.

Example 1 Identification of Olfactory Receptor Responsive to Sotron 1) Cloning of Human Olfactory Receptor Gene Human olfactory receptor is based on sequence information registered in GenBank and human genomic DNA female (G1521: Promega) as a template. The PCR method was used for cloning. Each gene amplified by the PCR method is incorporated into the pENTR vector (Invitrogen) according to the manual, and the NotI and AscI created downstream of the Flag-Rho tag sequence on the pME18S vector using the NotI and AscI sites present on the pENTR vector. Recomposed to the site.

2) Preparation of pME18S-human RTP1S vector A gene encoding a human receptor transport protein, RTP1S (GenBank GI: 50234917) was incorporated into the EcoRI and XhoI sites of the pME18S vector.

3) Preparation of olfactory receptor-expressing cells HEK293 cells each expressing 400 types of human olfactory receptors were prepared. A reaction solution having the composition shown in Table 2 was prepared and allowed to stand in a clean bench for 15 minutes, and then added to each well of a 96-well plate (BD). Next, 100 μL of HEK293 cells (3 × 10 ⁵ cells / cm ² ) were seeded in each well and cultured in an incubator maintained at 37 ° C. and 5% CO ₂ for 24 hours.

4) Luciferase assay The olfactory receptor expressed in HEK293 cells increases the amount of intracellular cAMP by coupling with intracellular Gαs and activating adenylate cyclase. For the measurement of the sotron response in this study, a luciferase reporter gene assay was used which monitors the increase in intracellular cAMP level as a luminescence value derived from the firefly luciferase gene (fluc2P-CRE-hygro). In addition, a renilla luciferase gene fused to the downstream of the CMV promoter (hRluc-CMV) was introduced at the same time, and used as an internal standard for correcting errors in gene transfer efficiency and cell number.
The medium was removed from the culture prepared in 3) above, and 75 μL of a solution containing sotron (1 mM) prepared in CD293 medium (Invitrogen) was added. The cells were cultured for 2.5 hours in a CO ₂ incubator to fully express the luciferase gene in the cells. The luciferase activity was measured using a Dual-Glo ^™ luciferase assay system (Promega) according to the operation manual of the product. A value obtained by dividing the luminescence value derived from firefly luciferase induced by sotron stimulation by the luminescence value in cells not subjected to sotron stimulation was calculated as a fold increase and used as an index of response intensity.

5) Results As a result of measuring responses to sotron (1 mM) for 400 kinds of olfactory receptors in this example, it has already been reported that they respond to sotron (Non-patent Document 1). A response was shown. On the other hand, other olfactory receptors did not respond to sotron (FIG. 1).

Example 2 OR8D1 concentration-dependent response to sotron In the same manner as in Example 1, the olfactory receptor OR8D1 (GenBank GI: 50879281, SEQ ID NO: 2) was expressed in HEK293 cells together with human RTP1S, and various concentrations of sotron ( 0, 3, 10, 30, 100, 300, 1000, and 3000 μM) were examined for concentration dependence. As a result, OR8D1 showed a concentration-dependent response to sotron (FIG. 2).

Example 3 Identification of OR8D1 antagonists 228 test substances were examined for antagonist activity of the olfactory receptor OR8D1 against the sotron response.
In the same procedure as in Example 2, sotron (100 μM) and test substance (100 μM) were added to HEK293 cells expressing olfactory receptor OR8D1, and the response of the olfactory receptor was measured. Response changes were evaluated.
The inhibition rate of the receptor response by the test substance was calculated as follows. A luminescence value (X) derived from firefly luciferase induced by stimulation with sotron alone is subtracted by a luminescence value (Y) in a cell in which the same receptor is introduced but not subjected to sotron stimulation, and the receptor is stimulated by sotron alone. Activity (XY) was determined. Similarly, the luminescence value (Z) by stimulation with a mixture of sotron and test substance is subtracted by the luminescence value (Y) in cells not subjected to sotron stimulation, and the receptor activity in the presence of the test substance (ZY ) Using the following formula, calculate the rate of decrease in receptor activity (ZY) in the presence of the test substance relative to receptor activity (XY) by sotron alone stimulation, and calculate the receptor response inhibition rate by the test substance. Asked. In the measurement, independent experiments were performed a plurality of times in duplicate, and an average value of each experiment was obtained.

Inhibition rate (%) = {1- (ZY) / (XY)} × 100

As a result, 10 kinds of test substances showed an inhibition rate of OR8D1 with respect to the sotron response of 40% or more (suppression of response to 60% or less), and showed an antagonist activity against OR8D1 (Table 3).

Example 4 Evaluation of Sotron Odor Suppressing Ability of OR8D1 Antagonist The sotron odor suppressing ability of a test substance having antagonist activity against OR8D1 identified in Example 3 was confirmed by a sensory test.
A sample containing 0.5 μL of a fragrance was added to 0.5 g of a dough containing sotron (1%), and the strength of the sotron odor with respect to the dough not fragranced was evaluated. The sensory evaluation test was conducted by three panelists, and the evaluation was made with 1 when the sotron odor was strongly felt and 5 when the sotron odor was not felt at all.
As a result, all 10 types of test substances that were shown to suppress the OR8D1 sotron response in Example 3 suppressed the sotron odor (FIG. 3).

Claims (1)

An inhibitor of odor caused by sotron, comprising at least one selected from the group consisting of the following compounds as an active ingredient:
1- (2,3,4,7,8,8a-hexahydro-3,6,8,8-tetramethyl-1H-3a, 7-methanoazulen-5-yl) -ethanone;
Oxacyclohexadecen-2-one;
Citral;
ω-6-hexadecenelactone;
2-benzylideneheptanal;
3- (4-tert-butylphenyl) propanal;
(5E) -3-methylcyclopentadec-5-en-1-one;
1- (5,6,7,8-tetrahydro-3,5,5,6,8,8-hexamethyl-2-naphthalenyl) -ethanone;
7-acetyl-1,2,3,4,5,6,7,8-octahydro-1,1,6,7-tetramethyl-naphthalene;
Muscon.